Aiming for the management of the small hive beetle, Aethina tumida, using relative humidity and diatomaceous earth

Small hive beetles (SHBs) are a global pest of European honeybee colonies. In the laboratory, the survival of adult SHBs was evaluated in relation to relative humidity (RH = 56, 64, 73, 82 and 96 %) and treatment with diatomaceous earth (DE) across 4 days. Low RH reduced survival. The application of DE reduced survival in addition to RH. Adults treated with corn flour (control) showed no difference in survival from untreated beetles. Scanning electron microscopy images showed no scarification of adult beetle cuticle after exposure to DE; therefore, water loss is likely facilitated through non-abrasive means such as the adsorption of cuticular lipids. The data agree with the hypothesis that DE causes mortality through water loss from treated insects. Egress, ingress, mortality and the egg-laying behaviours of beetles were observed in relation to a popular in-hive trench trap with and without the addition of DE. Traps filled with DE resulted in 100 % mortality of beetles compared with 8.6 % mortality when no DE was present. A simple method for visually determining beetle sex was used and documented.

Heat shock response in mulberry silkworm races with different thermotolerances

The thermal sensitivity and heat shock response of the different races of the mulberry silkworm Bombyx mori have been analysed. The multivoltine race, strains C. Nichi and Pure Mysore showed better survival rates than the bivoltine race, strain NB4D2 exposed to 41 degrees C and above. In general, the fifth instar larvae and the pupae exhibited maximum tolerance compared to the early larval instars, adult moths or the eggs. Exposure up to 39 degrees C for 1 or 2 h was tolerated equally whereas temperatures above 43 degrees C proved to be lethal for all. Treatment of larvae at 41 degrees C for Ih resulted in a variety of physiological alterations including increased heart beat rates, differential haemocyte counts, enlargement of granulocytes and the presence of additional protein species in the tissues and haemolymph. The appearance of a 93 kDa protein in the haemolymph, fat bodies and cuticle, following the heat shocking of larvae in vivo was a characteristic feature in all the three strains examined although the kinetics of their appearance itself was different. In haemolymph, the protein appeared immediately in response to heat shock in C. Nichi reaching the maximal levels in 2-4 h whereas its presence was noticeable only after 2-4 h recovery time in Pure Mysore and bivoltine races. The fat body from both C. Nichi and NB4D2 showed the presence of 93 kDa, 89 kDa and 70 kDa proteins on heat shock. The haemocytes, on the other hand, expressed only a 70 kDa protein consequent to heat shock. The 93 kDa protein in the haemolymph, therefore could have arisen from some other tissue, possibly the fat body. The 93 kDa protein was detected after heat shock in pupae and adult moths as well, although the presence of an additional (56 kDa) protein was also apparent in the adults. The presence of 46 kDa and 28 kDa bands in addition to the 93 kDa band in the cuticular proteins immediately following heat shock was clearly discernible. The 70 kDa band did not show much changes in the cuticular proteins on heat shock. In contrast to the changes in protein profiles seen in tissues and haemolymph following heat shock in vivo, the heat treatment of isolated fat body or haemolymph in vitro resulted in protein degradation.

Effect of SDS on human hair: Study on the molecular structure and morphology

This paper presents a model study to understand the effect of surfactants on the physicochemical properties of human hair. FT-IR ATR spectroscopy has been employed to understand the chemical changes induced by sodium dodecyl sulfate (SDS) on human scalp hair. In particular, the SDS induced changes in the secondary structure of protein present in the outer protective layer of hair, i.e. cuticle, have been investigated. Conformational changes in the secondary structure of protein were studied by curve fitting of the amide I band after every phase of SDS treatment. It has been found that SDS brings rearrangements in the protein backbone conformations by transforming beta-sheet structure to random coil and beta-turn. Additionally, AFM and SEM studies were carried out to understand the morphological changes induced on the hair surface. SEM and AFM images demonstrated the rupture and partial erosion of cuticle sublayers.

Colored polydimethylsiloxane micropillar arrays for high throughput measurements of forces applied by genetic model organisms

Measuring forces applied by multi-cellular organisms is valuable in investigating biomechanics of their locomotion. Several technologies have been developed to measure such forces, for example, strain gauges, micro-machined sensors, and calibrated cantilevers. We introduce an innovative combination of techniques as a high throughput screening tool to assess forces applied by multiple genetic model organisms. First, we fabricated colored Polydimethylsiloxane (PDMS) micropillars where the color enhances contrast making it easier to detect and track pillar displacement driven by the organism. Second, we developed a semiautomated graphical user interface to analyze the images for pillar displacement, thus reducing the analysis time for each animal to minutes. The addition of color reduced the Young's modulus of PDMS. Therefore, the dye-PDMS composite was characterized using Yeoh's hyperelastic model and the pillars were calibrated using a silicon based force sensor. We used our device to measure forces exerted by wild type and mutant Caenorhabditis elegans moving on an agarose surface. Wild type C. elegans exert an average force of similar to 1 mu N on an individual pillar and a total average force of similar to 7.68 mu N. We show that the middle of C. elegans exerts more force than its extremities. We find that C. elegans mutants with defective body wall muscles apply significantly lower force on individual pillars, while mutants defective in sensing externally applied mechanical forces still apply the same average force per pillar compared to wild type animals. Average forces applied per pillar are independent of the length, diameter, or cuticle stiffness of the animal. We also used the device to measure, for the first time, forces applied by Drosophila melanogaster larvae. Peristaltic waves occurred at 0.4Hz applying an average force of similar to 1.58 mu N on a single pillar. Our colored microfluidic device along with its displacement tracking software allows us to measure forces applied by multiple model organisms that crawl or slither to travel through their environment. (C) 2015 AIP Publishing LLC.

Experimental study on the microstructure and nanomechanical properties of the wing membrane of dragonfly

Detailed investigations on the microstructure and the mechanical properties of the wing membrane of the dragonfly are carried out. It is found that in the direction of the thickness the membrane was divided into three layers rather than a single entity as traditionally considered, and on the surfaces the membrane displays a random distribution rough microstructure that is composed of numerous nanometer scale columns coated by the cuticle wax secreted. The characteristics of the surface structure are measured and described. The mechanical properties of the membranes taken separately from the wings of live and dead dragonflies are investigated by the nanoindentation technique. The Young's moduli obtained here are approximately two times greater than the previous result, and the reasons that yield the difference are discussed.

Microstructure and nanomechanical properties of the wing membrane of dragonfly

Detailed investigations on the microstructure and the mechanical properties of the wing membrane of the dragonfly were carried out. It was found that in the direction of the thickness the membrane was divided into three layers rather than as traditionally considered as a single entity, and on the surfaces the membrane displayed a random distribution rough microstructure that was composed of numerous nanometer scale columns coated by the cuticle wax secreted. The characteristics of the surfaces were accurately measured and a statistical radial distribution function of the columns was presented to describe the structural properties of the surfaces. Based on the surface microstructure, the mechanical properties of the membranes taken separately from the wings of living and dead dragonflies were investigated by the nanoindentation technique. The Young's moduli obtained here are approximately two times greater than the previous result, and the reasons that yield the difference are discussed. (C) 2007 Elsevier B.V. All rights reserved.

Plankton associated with medusae of the freshwater jellyfish Craspedacusta sowerbyi (Lankester) in a Thames backwater

Samples of plankton were taken from Broom Water in August 1997 after a sighting of medusae on 11th August. Broom Water is about 8 m wide, and extends 250 m from the main channel of the Thames, above the weir at Teddington. On 11th August medusae were so abundant that it was possible to collect 20 in ten minutes. They were rising to the surface in bright sunlight, then sinking slowly down through the water. Examination of a medusa's tentacles under a microscope revealed the presence of a commensal protozoan, a ciliate Trichodina pediculus. Over 20 species of phytoplankton were found in Broom Water. Most of the species are common and widespread, but it was a surprise to find Errerella bornhemiensis with its characteristic pyramidal colonies, which is a relatively rare species. Zooplankters in Broom Water consisted of Rotifera and Crustacea. Zooplankton is the main food of Craspedacusta and it was found that the crustaceans but not the rotifers did undergo significant changes during the period 11-19th August. The major changes were a big increase in the percentage of cyclopoids, and a marked decrease in Bosmina. This could be because the delicate cuticle of Bosmina is much more susceptible to the stinging cells of the medusae compared with the tougher exoskeleton of the cyclopoid.

I. Studies on the activation of Drosophila phenol oxidase. II. Studies on serum insulin in normal subjects and diabetics

Part I

Phenol oxidase is the enzyme responsible for hardening and pigmentation of the insect cuticle. In Drosophila, phenol oxidase is a latent enzyme. Enzyme activity is produced by the interaction of a number of protein components. A minimal activation scheme consisting of six protein components, designated Pre S, S activator, S, P. P' and Ʌ₁ is described. Quantitative assays have been developed for the S activator, S, P and P' proteins and these components have been partially purified. Experiments describing the interactions of the six components have been conducted and a model for the activation of phenol oxidase in a minimal system is proposed. Possible mechanisms of the reactions between the constituents of the activating system and potential regulatory mechanisms involved in phenol oxidase production and function are discussed.

Part II

A method has been developed for the partial purification of insulin from human serum. A procedure for the determination of the electrophoretic mobility of serum insulin on polyacrylamide gels is described. An electrophoretic analysis of insulin isolated from a normal subject is reported and in addition to a major band, the existence of a number of minor bands of immunoreactive insulin is described. A comparison of the electrophoretic patterns of insulin isolated from normal and diabetic subjects was carried out and indications that differences between them may occur are reported.

Desarrollo larval de dos especies de Crustáceos Decápodos en el laboratorio, Pachycheles haigae Rodrigues Da Costa (Porcellanidae) y Chasmagnathus granulata Dana (Grapsidae)

The present paper is concerned with studies on the larval development of two species of crustacean Decapoda: Pachyceles haigae Rodrigues Da Costa, 1960 family Porcellanidae and Chasmagnathus granulata Dana, 1851, family Grapsidae. One preazoea, two zoeae with a total of five sub-stages, a and b, and the second zoea had sub-stages a, b and c. The greatest changes are observed in the transition from the first to the second zoea and from this to the megalopa. During the sub-stages, there occured minor changes, and a net moulting of the cuticle is not observed. The differences in these sub-stages are marked by changes in the number and shape of the appendages.

Palinologia e taxonomia dos gêneros Elephantopus L., Orthopappus Gleason e Pseudelephantopus Rohr. (Subtribo Elephantopinae, Vernonieae- Asteraceae) ocorrentes no Brasil

A subtribo Elephantopinae tem sido alvo de poucos estudos taxonômicos e palinológicos, necessitando de uma reavaliação em seus limites. Assim, o presente estudo teve como proposta atualizar e ampliar o conhecimento de 13 espécies de Elephantopinae ocorrentes no Brasil através de um acurado estudo palinológico, da anatomia foliar e taxonômico, promovendo a delimitação das espécies dos gêneros Elephantopus (10 espécies) Orthopappus (uma espécie) e Pseudelephantopus (duas espécies)além de oferecer subsídios para posteriores análises filogenéticas. O material botânico utilizado foi obtido através de exsicatas depositadas nos herbários brasileiros e de material coletado em campo. Os grãos de pólen foram tratados pelo método acetolítico, sendo posteriormente mensurados, descritos, e analisados sob microscopia de luz e eletrônica de varredura. As eletromicrografias foram obtidas de grãos de pólen não acetolisados, as análises taxonômicas baseiam-se na metodologia clássica. A anatomia foliar deu-se através da metodologia usual. Os resultados mostram grãos de pólen médios, 3-porados, de exina equinolofada com malhas poligonais organizadas ou não em Elephantopus e Pseudelephantopus, podendo apresentar interrupção na malha poral em Elephantopus. Em Orthopapus os grãos de pólen são 3-colporados, de sexina subequinolofada. A anatomia foliar possibilitou a separação de E. hirtiflorus e E. riparius através da forma ou contorno da nervura principal (planoconvexo), nas demais espécies o contorno é biconvexo. As espécies apresentaram tricomas capitados bisseriados com 8 células, tricomas filamentosos 3-4 celulares, bem como filamentosos de célula distal globosa ou ovóide em E. biflorus, E. micropappus, E. tomentous e Orthopappus angustifolius. Registrou-se variação na ornamentação da cutícula podendo ser lisa ou estriada entre as espécies, bem como a presença de substâncias pécticas na epiderme e drusas nos parênquimas. Em relação aos microcaracteres florais, foram considerados de importância diagnóstica os lóbulos da corola, que variaram entre glandular, glabro e penicilado, este ultimo apenas em E. hirtiflorus, os ápices das anteras que variaram entre obtuso, retuso em E. hirtiflorus e apiculado, lancelolado em E. mollis e E. racemosus, a base da antera variou entre sagitada, caudada lisa em E. riparius e E. tomentosus e caudada em E. riparius. Os macrocaracteres que apresentaram maior valor taxonômico foram a cipsela e o papus, a organização das coflorescências, o número de flores por capítulo e limbo foliar. Neste trabalho aceitou-se a segregação dos três gêneros, com base nos caracteres analisados

浙江嵊州早上新世植物群重建和古气候定量研究

上新世时期是地球环境发展变化过程中由温暖的中新世向冰期-间冰期交替出现的更新世的过渡时期，研究上新世的气候有助于研究现代全球气候变化趋势。我国上新世植物群研究不多，尤其缺乏早上新世植物群的研究。在研究化石植物群的过程中，叶角质层的信息通常被忽视。迄今，我国古气候的定量研究主要集中在山东山旺和云南地区，研究的主要是中新世和晚上新世的古气候，早上新世气候的定量研究国内外都未见报道。 本论文以采自浙江中部嵊州地区硅藻土中的植物叶和果实为材料，通过光学显微镜、荧光显微镜和扫描电镜观察，对提取的化石叶角质层和叶结构进行研究，对比研究现代相关类群植物的叶片，确定了化石植物类群计15科24属（或亚科）34种。同时参照现代植物分布，恢复了早上新世嵊州地区古植被情况，进一步运用共存分析（Co-existence Approch）定量重建了当时的古气候，得到古年均温、古年较差和古年降水量等7个参数。 研究表明，叶结构和叶角质层可以应用于化石植物类群的鉴定。光学显微镜和荧光显微镜下可以获得的信息有：气孔、表皮细胞形状及大小、表皮细胞垂周壁、分泌结构、表皮毛或毛基等。扫描电镜观察的是叶表皮细胞表面的情况，如细胞外壁是否被蜡质以及蜡质的交结情况，气孔外壁是否增厚呈环等。 经化石植物类群鉴定，早上新世（约4Ma）嵊州地区植被属亚热带常绿阔叶林。推测：该地区当时存在一个较大的湖泊，湖泊边缘水中生长有菱角，湖泊周边可能有海拔超过1400m的山地。 常绿阔叶林主要成分是壳斗科植物，其中青冈属植物可能是建群植物。自低海拔到高海拔地区都有分布的是：栎属、栗属、桤木属植物，青杨梅。低海拔地区（<600m）分布的植物有：海南锥，樟。中低海拔地区（300-1000m）分布着：柯属、黄杨属植物，水青冈，海南油丹，江南油杉，福建柏。中高海拔地区（>700m）分布有：高山栎、黄肉楠属植物，米心水青冈，榉树，建始槭，天台鹅耳枥和昌化鹅耳枥。除了上述类群，林中还散布着鼠李属、杜鹃花属和冬青属的植物，豆科崖豆藤属植物则缠绕一些较大乔木生长。另外，在一些土壤、有机质易堆积的洼地生长有竹林，山地贫瘠的地方生长着柏属植物。 嵊州早上新世化石植物群反映的植被同浙江地区现代植被相似。第三纪浙江地区从针叶林或阔叶树为主的针-阔叶混交林过渡到常绿-落叶阔叶林，再发展为常绿阔叶林。 依据23个植物类群分析获得7个古气候参数：年均温17.7-21.4°C，最热月均温22.2-27°C，最冷月均温8.4-13.4°C，年较差12.1-15.3°C，年降水量1136-1869.9mm，最大月降水量211.8-283.3mm，最小月降水量20.3-36.8mm。 对比现代嵊州地区气候参数，早上新世嵊州地区年降水量略高于现代，年均温比现代高1.3-5°C，年较差为12.1-15.3°C。早上新世期嵊州地区四季气温比现代平稳。

杉科植物的表皮结构研究及古环境重建

全球古近纪和新近纪气候波动明显，很多科学家对古近纪和新近纪生物演化和气候演变规律的定量研究给予了相当的重视。杉科植物有长期发展的历史、少数的现存种和丰富的化石记录，成为指示古环境的理想植物之一。利用杉科植物重建古环境，首先要对化石植物进行正确的分类鉴定。杉科各属枝叶形态特征变异幅度较大，且枝叶排列和形态特征近似，有时各属之间存在交叉特征。杉科化石标本通常仅保存枝叶形式，且多个属的化石标本经常发现于同一地层，因而分类鉴定比较困难。植物的叶表皮结构是压型植物化石细胞信息的重要来源，是属种分类鉴定重要依据之一。本文在以往研究的基础上，完整分析了杉科9属现生植物的表皮特征，建立了杉科化石植物分类鉴定的现生植物的表皮特征参照系。 杉科植物的叶多为条形、钻形、鳞形或披针形，同种植物有1种、2种甚至3种叶型。其中水杉属的叶交互对生，其它属的叶螺旋状互生。水杉属的多数表皮细胞垂周壁明显弯曲，落羽杉属和杉木属有时微呈波状，其他属的表皮细胞垂周壁直。多数植物的叶片近轴面和远轴面的气孔数量和分布不同。一般来说，条形叶和披针形叶的远轴面气孔分布状况和气孔数量稳定，远轴面的中部最稳定。条形叶远轴面的气孔分布于中脉两侧，形成纵向的气孔带。条形叶近轴面气孔分布状况和气孔数量变化大，近轴面的气孔数量有时与远轴面近似，但多数情况下比远轴面少，有时整个叶片的近轴面仅少数几个气孔或没有气孔分布。钻形叶的近轴面和远轴面的气孔数量近似，或叶片近轴面的气孔数量比远轴面的气孔数量多，气孔分布范围也比远轴面气孔分布范围广。气孔椭圆形，落羽杉属植物和水松鳞形叶的气孔长轴方向与叶片长轴垂直或斜向排列，柳杉属植物的气孔多斜向排列，水松的条形叶和条状钻形叶的气孔多平行向排列。落羽杉属和柳杉属以外的杉科植物的气孔长轴多数与叶片长轴平行。在扫描电子显微镜下的密叶杉属植物叶片角质层的内表面，副卫细胞和表皮细胞的垂周壁与叶表面的角度一致，且副卫细胞的形态与表皮细胞类似，这种特殊气孔器被称为A-型气孔器。杉科植物中水杉属、落羽杉属和杉木属的气孔器近似密叶杉的这种A-型气孔器。台湾杉属、柳杉属、水松属、红杉属和巨杉属植物的气孔器与这种A-型气孔器不同，这几属的副卫细胞垂周壁表皮细胞的垂周壁方向不同，且副卫细胞的形态与表皮细胞的形态明显不同。因而气孔所有副卫细胞组成呈明显的圆盘状，保卫细胞在盘子的中央。综合分析植物的枝叶形态和表皮特征可以区分杉科各属。 本文研究了采自抚顺始新世的杉科植物标本，经枝叶特征、表皮结构和球果鳞片等特征分析为水松属的欧洲水松。论文分析了中国东北地区古温度定量研究结果与杉科植物的相关性。抚顺植物群发现有化石水杉M. occidentalis、中华红杉和欧洲水松，根据这三种化石植物的年均温度和年降水量，通过共存分析，推测抚顺始新世的年均温为12.1-17℃，年降水量为1199.6-2231mm。

不同贮藏条件下果实角质层成分及其与耐贮性关系研究

角质层是果实抵御外界环境胁迫的一个屏障，既能调节果实自身的生理活动，也能增加对病原菌入侵的抵抗力，在果实贮藏保鲜中具有重要的作用。本文利用傅里叶变换红外光谱检测（Fourier-transform-infrared-spectroscopy）法，重点研究了不同贮藏条件下果实角质层和果肉细胞壁成分变化，及对果实品质和贮藏性的影响，为进一步阐明角质层在果实贮藏保鲜中的作用提供依据。研究内容包括：（1）壶瓶枣果实在冷藏和气调下软化率、品质、角质层和果肉细胞壁成分变化；（2）即时冷藏和延迟冷藏下，桃果实好果率、品质、角质层和果肉细胞壁成分变化；（3）采前喷施氯化钙或油菜素内酯对甜樱桃单果重、品质、果肉细胞壁结构以及耐贮性的影响。 试验结果表明：（1）与冷藏（-1±1 ºC）相比，气调贮藏（10% O2 + 0% CO2， -1±1 ºC）能够显著降低壶瓶枣软化率，更好地保持果实的硬度、可溶性固形物（SSC）和可滴定酸（TA）含量以及较高含量的果肉细胞壁物质和较低含量的角质层物质。（2）与延迟冷藏（25 ºC，48 h后转入0 ºC）相比，即时冷藏使“八月脆”桃果实能保持较高的果实硬度和好果率，显著减慢TA含量下降，能保持较高含量的果肉细胞壁物质和角质层物质，但对SSC和Vc含量没有显著的影响。（3）与对照相比，采前喷施CaCl2（1%，m/v）能够增加“红灯”（6.94%）的单果重，对甜樱桃果实的品质指标（硬度、SSC、TA）和细胞壁结构没有明显的影响。（4）与对照相比，采前喷施0.15 mg•L-1油菜素内酯能增加“红灯”（3.68%）和“大紫”（8.61%）的单果重，降低“红灯”果实的自然腐烂率，并不影响果实的品质指标（硬度、SSC、TA）。 这些研究结果说明：（1）与冷藏（-1±1 ºC）相比，气调贮藏（10% O2 + 0% CO2， -1±1 ºC）更利于壶瓶枣果实贮藏保鲜；（2）与延迟冷藏（25 ºC，48 h后转入0 ºC）相比，即时冷藏（0 ºC）更利于“八月脆”桃果实贮藏保鲜；（3）气调贮藏和即时冷藏通过调节果实角质层和细胞壁代谢等途径发挥作用。气调贮藏会降低壶瓶枣果实角质层物质含量，增强其透气性，减少壶瓶枣酒软发生；但即时冷藏会延缓“八月脆”桃果实角质层降解，维持角质层物质较高的含量以及结构完整性，以充分发挥角质层的保护作用，减缓果实的软化进程，维持果实硬度和品质；（4）采前适当浓度的钙或油菜素内酯处理对增加甜樱桃单果重，降低自然腐烂率有一定的作用，但在不同品种中的作用效果有差异。

Artificial insemination in penaeid shrimps

Artificial Insemination (AI) is a tool for genetic manipulation in the shrimp stocks. It is seen as one of the means for propagating shrimp culture to new areas by controlled reproduction. Attempts at artificial insemination in the dominant closed-thelycum penaeid shrimps species of the area viz. Metapenaeus affinis and Metapenaeus brevicornis were induced in wild adult stocks collected off Mumbai coast. Female specimens were subjected to unilateral eyestalk ablation by pinching so as to induce moulting and maturation. AI was performed two days after moulting on these females when the cuticle was still soft and flexible. Moulting also ensured rejection of initial spermatophores, if present. Response of males to electrical stimulation for spermatophore expulsion was spontaneous. Use of tissue glue for spermatophore retention was found to be unnecessary. Latency period ranged between10-16 days, while spawning occurred within 10-12 days of spermatophore transfer. Three partial spawning were recorded viz., two in Metapenaeus affinis and one in Metapenaeus brevicornis with an average spawning and hatching rates of 30% and 72.3% respectively. Average survival from first nauplius (N1) to one-day old post-larva (PLI) was a meager 3.43%. Use of AI in genetic manipulation of shrimp stocks for aquacultural purposes is indicated.

In vivo dynamics of the internal fibrous structure in smooth adhesive pads of insects.

Many insects with smooth adhesive pads can rapidly enlarge their contact area by centripetal pulls on the legs, allowing them to cope with sudden mechanical perturbations such as gusts of wind or raindrops. The short time scale of this reaction excludes any neuromuscular control; it is thus more likely to be caused by mechanical properties of the pad's specialized cuticle. This soft cuticle contains numerous branched fibrils oriented almost perpendicularly to the surface. Assuming a fixed volume of the water-filled cuticle, we hypothesized that pulls could decrease the fibril angle, thereby helping the contact area to expand laterally and longitudinally. Three-dimensional fluorescence microscopy on the cuticle of smooth stick insect pads confirmed that pulls significantly reduced the fibril angle. However, the fibril angle variation appeared insufficient to explain the observed increase in contact area. Direct strain measurements in the contact zone demonstrated that pulls not only expand the cuticle laterally, but also add new contact area at the pad's outer edge.